KR101729687B1 - Method for manufacturing suprerparamagnetic nanocomposite and suprerparamagnetic nanocomposite manufactured by the method - Google Patents

Abstract

The present invention relates to a method for manufacturing a superparamagnetic nanocomposite and a superparamagnetic nanocomposite manufactured using the same and, more specifically, relates to a method for manufacturing a superparamagnetic nanocomposite, which can be used for magnetic isolation in the detection of a biological target material, and a superparamagnetic nanocomposite manufactured using the same. According to the present invention, the method for manufacturing a superparamagnetic nanocomposite can mass-produce superparamagnetic nanocomposites with excellent characteristics including a uniform size and a uniform particle size distribution, high aqueous solution dispersibility, and maintained superparamagnetism and a high degree of magnetization, at a high yield and at a high speed without complicated processes as compared to the conventional method for manufacturing a magnetic nanoparticle for magnetic isolation. The superparamagnetic nanocomposite manufactured by the method maintains superparamagnetism and has a high degree of magnetization. The method comprises the following steps of: mixing an iron precursor, a solvent, a stabilizer and a reducing agent; synthesizing a nanoclustered superparamagnetic nanocomposite by hydrothermally synthesizing the mixed solution in the mixing step at a temperature of 150 to 300C and a pressure of 1.5 to 10 bar; and isolating the synthesized superparamagnetic nanocomposite.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing a super magnetic nanocomposite and a super magnetic nanocomposite prepared by using the same,

TECHNICAL FIELD The present invention relates to a method for producing a super-magnetic nanocomposite and a super-magnetic nanocomposite fabricated using the same, and more particularly, to a method for manufacturing a super-magnetic nanocomposite applicable to the detection of a biomolecular target material, Lt; / RTI >

Development of methods for highly sensitive detection and quantification of biomolecules such as target biomarkers in the fields of medicine and life sciences such as diagnosis of diseases and development of new drugs is considered to be very important. Most commonly, a binding assay based on an antigen-antibody immune reaction, DNA hybridization or receptor reaction is widely used depending on the target substance And the presence of a target molecule is determined by a signal transducer that converts a binding reaction to a target molecule into a measurable signal.

  The magnetic nanoparticle mediated isolation technique using magnetic force in this coupled reaction assay can be obtained by concentrating only the target biomolecule from a suspension solution containing various impurities or non-target substances (positive isolation), or Negative isolation of non-target molecules provides advantages such as simplification of the assay, ease of processing, sensitivity, specificity improvement, high throughput screening, and scalability.

Magnetic nanoparticle-based separation technology involves ligating a ligand material that specifically binds to a target molecule to particles, recognizing and binding target molecules of the ligand material in the mixed solution, and separating the magnetic particles using external magnetic force do. As a target molecule sensing platform, the magnetic particles should (i) be capable of minimizing non-specific adsorption from a variety of non-specific materials in the suspension solution, (ii) capable of minimizing colloid particle stability from a variety of biochemical environments, (Iii) it should be easy to bond the surface of various functional groups. In order to have a non-fouling excellent bio-interface, it is preferable to have hydrophilicity and neutral and have a characteristic including hydrogen bond acceptors. For this purpose, pegylation, that is, coating of poly (ethylene glycol), a biocompatible polymer on the surface of particles, is one of the most successful methods of designing nonadhesive living surface nanoparticles to date It is considered. The precisely adsorbed PEG layer satisfies the conditions listed above, reduces nonspecific adsorption of the particles, and increases stability.

The present inventors have provided a method for producing a super-magnetic nanocomposite, that is, a super-magnetic iron oxide nanocomposite that can be used for the purpose of magnetic separation for the detection of a biomolecular target, The present invention relates to a process for producing a super-magnetic nanocomposite having excellent properties, which has a uniform size and particle size distribution, a high aqueous dispersion dispersibility, a super magnetism and a high magnetization without a complicated process at a yield. Completed.

Effective PEGylation of iron oxide nanoparticles for in vivo cancer imaging. Advanced Functional Materials, 21 (8), 1498-1504, 2011.

An object of the present invention is to provide a super-magnetic nanocomposite having excellent properties of dispersing a high aqueous solution, having a uniform size and a particle size distribution at a high speed without complicated processes at a high yield, In a large amount, and to provide a super-magnetic nanocomposite produced by the above method.

The present invention relates to a process for preparing iron precursors, comprising the steps of: mixing an iron precursor, a solvent, a stabilizer and a reducing agent; The mixed solution is heated at a temperature of 150 to 300 ° C, preferably 200 to 240 ° C, more preferably 200 ° C, and a temperature of 1.5 to 10 bar, preferably 1.5 to 6 bar. , More preferably 1.5 to 2.5 bar, to synthesize a nanoclustered super-magnetic nanocomposite; And separating the synthesized super-magnetic nanocomposite. The present invention also provides a method for producing a super-magnetic nanocomposite.

The method for preparing a super-magnetic nanocomposite according to the present invention may further comprise washing the separated super-magnetic nanocomposite with a polar solvent.

In the method for producing a superparamagnetic nanocomposite according to the present invention, the iron precursor is selected from the group consisting of iron chloride hexahydrate (FeCl ₃ .6H ₂ O), iron chloride (II), iron chloride heptahydrate (II) (Fe (NO ₃ ) ₃ .9H ₂ O). Preferably, the iron precursor may be selected from the group consisting of iron chloride hexahydrate (FeCl ₃ .6H ₂ O), iron (II) chloride, iron (II) chloride and iron (III) chloride, (FeCl ₃ .6H ₂ O).

In the method for producing a super-magnetic nanocomposite according to the present invention, the solvent may be selected from the group consisting of ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, dipropylene glycol and glycerol. Preferably, the solvent may be ethylene glycol.

In the method for preparing a super-magnetic nanocomposite according to the present invention, the stabilizer may be a compound having a carboxyl group.

In the method of preparing a super-magnetic nanocomposite according to the present invention, the stabilizer is at least one selected from the group consisting of trisodium citrate dihydrate (HOC (COONa) (CH ₂ COONa) ₂ .2H ₂ O; C ₆ H ₅ Na ₃ O ₇ And a dicarboxylic poly (ethylene glycol) having a molecular weight of 500 to 50,000, preferably a molecular weight of 2000 to 8000, and more preferably a molecular weight of 2000.

In the method for preparing a super-magnetic nanocomposite according to the present invention, the reducing agent may be selected from the group consisting of sodium acetate, sodium acrylate, urea, sodium formate, and ammonium acetate. Preferably, the reducing agent may be sodium acetate.

In the method of preparing the super-magnetic nanocomposite according to the present invention, the iron precursor and the solvent may be mixed in a molar ratio of 1:10 to 1: 300, preferably 1:40 to 1: 200.

In the method of preparing the super-magnetic nanocomposite according to the present invention, the iron precursor and the stabilizer may be mixed in a molar ratio of 1: 0.0000013 to 1: 1, preferably 1: 0.0000013 to 1: 0.8 .

In the method for producing a superpowder nanocomposite according to the present invention, the iron precursor and the reducing agent are mixed in a molar ratio of 1: 1 to 1:20, preferably 1: 3 to 1:15, more preferably 1: : 7 to 1: 15.

In the method for producing a superpowder magnetic nanocomposite according to the present invention, the polar solvent is preferably selected from the group consisting of ethanol, water, methanol, acetone, liquid ammonia, ethyl acetate, ether, tetrahydrofuran, potassium hydroxide, sodium hydroxide and dichloromethane Can be selected from the group.

In the method for producing a super-magnetic nanocomposite according to the present invention, the step of separating the synthesized super-magnetic nanocomposite may be separated by using a centrifugal separator or magnetism. The step of separating the synthesized super-magnetic nanocomposite may be performed by using a centrifuge or a method commonly used for separating magnetic nanoparticles using magnetism.

In the method of preparing a super-magnetic nanocomposite according to the present invention, the step of washing the separated super-magnetic nanocomposite with a polar solvent includes separating the super-magnetic nanocomposite separated in the step of separating the synthesized super- And washing with a solvent to remove impurities, so that the super-magnetic nanocomposite has high stability and uniform particle distribution. The polar solvent may be any one of ethanol, alcohol, liquid ammonia, acetone, methanol, chloroform, ethyl acetate, ether, tetrahydrofuran, potassium hydroxide, sodium hydroxide, dichloromethane and water. In the step of washing the separated super-magnetic nanocomposite with a polar solvent, it is preferable to wash the separated super-magnetic nanocomposite with a polar solvent three times. In this case, the washing may be performed from one time to several times, not limited to three times of washing, and all of the simple working variations on the number of times of washing are all within the scope of the present invention.

Meanwhile, the super-magnetic nanocomposite may be prepared without washing the separated super-magnetic nanocomposite with a polar solvent. However, in order to have high stability and uniform particle distribution as described above, It is preferable to carry out a step of washing the superficial magnetic nanocomposite with a polar solvent. The separation of the separated superparamagnetic nanocomposite with a polar solvent may be carried out by any of the conventional methods. At this time, the step of washing the separated super-magnetic nanocomposite with a polar solvent may be carried out by using a centrifugal separator, which is one of common methods. In the step of separating the synthesized super-magnetic nanocomposite, And it is within the scope of the present invention to complete the cleaning. This is because the step of separating the synthesized super magnetic nanocomposite may be divided into a first step and a second step, and the step of separating and washing may be performed together.

In the method of preparing a super-magnetic nanocomposite according to the present invention, the step of washing the separated super-magnetic nanocomposite with a polar solvent includes washing the separated super-magnetic nanocomposite with an ethanol solvent, And washing the super-pure magnetic nanocomposite washed with a water solvent. The step of washing with the ethanol solvent may be advantageous in terms of surface charge and the like by cleaning the solvent and the reducing agent with an ethanol solvent which is a polar solvent which is easy to dissolve. In addition, the step of washing the super-magnetic nanocomposite washed with the ethanol solvent using a water solvent may be advantageous in that deionized water can be dispersed in order to use it for the purpose of magnetic separation for detecting a biomolecular target substance .

In the method for preparing a super-magnetic nanocomposite according to the present invention, the super-magnetic nanocomposite can be controlled in its dispersibility in an aqueous solution by the carboxylate (COO ^- ) group of the stabilizer.

In the method for producing a super-magnetic nanocomposite according to the present invention, the super-magnetic nanocomposite may have a diameter of 100 nm to 450 nm. The super-magnetic nanocomposite may have a diameter of preferably 150 nm to 400 nm, more preferably 200 nm to 350 nm.

The present invention provides a super-magnetic nanocomposite produced by the method for producing the super-magnetic nanocomposite.

In the super-magnetic nanocomposite according to the present invention, the super-magnetic nanocomposite may have a diameter of 100 nm to 450 nm. The super-magnetic nanocomposite may have a diameter of preferably 150 nm to 400 nm, more preferably 200 nm to 350 nm.

In the super-magnetic nanocomposite according to the present invention, the super-magnetic nanocomposite includes magnetic nano-crystals having a diameter of more than 0 to 10 nm and stabilized by a carboxylate (COO ^- ) group, and a plurality of magnetic nano- May be aggregated and have a nanoclustered shape with a diameter of 100 nm to 450 nm and may have a hydrophilic property dispersed in an aqueous solution. The superparamagnetic nanocomposite may be in a nanoclustered form with a diameter of preferably 150 nm to 400 nm, more preferably 200 nm to 350 nm.

In the super-magnetic nanocomposite according to the present invention, the magnetic nano-crystal may be Fe ₃ O ₄ having a diameter of more than 0 and 10 nm or less.

The present invention has a diameter of less than 0 to 10nm carboxylate (COO ^-) groups of Fe ₃ surface is stabilized by O ₄ which comprises the magnetic nano-crystal, a plurality of magnetic nanoparticles is Crystal agglomeration is 100 nm in diameter to 450 nm nanoclustered nanocomposite having hydrophilicity dispersed in an aqueous solution. The superparamagnetic nanocomposite may be in a nanoclustered form with a diameter of preferably 150 nm to 400 nm, more preferably 200 nm to 350 nm.

The method for producing super magnetic nanocomposite according to the present invention has a higher speed and uniform size and particle size distribution than a conventional magnetic nanoparticle for magnetic separation at a higher yield without complicated processes, The superpowder magnetic nanocomposite having excellent magnetic properties can be produced in large quantities with excellent characteristics and excellent magnetization while retaining the superpowder magnetism and the superpowder magnetic nanocomposite produced by the above method has high magnetization , And can be used for the purpose of magnetic separation for biomolecular target detection.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic diagram showing a method for producing a super-magnetic nanocomposite (specifically, a super-magnetic iron oxide nanocomposite) in the form of magnetic nanoclusters according to Examples 1 and 2. FIG.
FIG. 2 shows the results of observation of the super-magnetic nanocomposites of Example 1 and Example 2 by scanning electron microscopy.
Fig. 3 shows the result of size measurement of the super-magnetic nanocomposite of Example 1. Fig.
Fig. 4 shows the results of measurement of the zeta potential of the super-magnetic nanocomposite of Example 1. Fig.
Fig. 5 shows the result of size measurement of the super-magnetic nanocomposite of Example 2. Fig.
Fig. 6 shows the results of measurement of the zeta potential of the super-magnetic nanocomposite of Example 2. Fig.
7 shows the results of measurement of the magnetic properties of the super-magnetic nanocomposite of Example 1. Fig.
8 shows the results of measurement of magnetic properties of the super-magnetic nanocomposite of Example 2. Fig.
FIG. 9 is a microscope image of a tube photograph taken before and after collection of erythrocytes, and a captured super-magnetic nanocomposite and supernatant.

Hereinafter, the present invention will be described in more detail using examples and the like. However, the following examples are provided for illustrating the present invention, and the present invention is not limited to the following examples, and various modifications and changes may be made thereto.

In the present invention, the super-magnetic nanocomposite is a single magnetic particle having a diameter of several nanometers (having a diameter of more than 0 to 10 nm), that is, a super-magnetic particle having a size of 100 to 450 nanometers in which magnetic nanocrystals are clustered in the form of clusters , Preferably in the range of 150 to 400 nanometers, more preferably in the range of 200 to 350 nanometers.

In the present invention, the normal temperature means 15 ° C to 25 ° C at which the operator can react most comfortably without raising or lowering the temperature, but the present invention is not limited thereto. That is, the temperature may be lower or higher depending on the surrounding environment and environment.

At this time, since the super-magnetism can be controlled by using magnetic force and can be re-dispersed when the magnetic force is lost, the super-magnetic nanocomposite can be used in various fields requiring magnetic nanoparticles having super magnetism.

Reference Example  1. Materials Preparation

Sodium citrate dihydrate (HOC (COONa) (CH ₂ COONa) ₂ .2H ₂ O; C (CH ₃ COONa) ₂ H ₂ O), iron (III) chloride hexahydrate (FeCl ₃ .6H ₂ O, ACS reagent, 97%, MW = 270.30) _{6 H 5 Na 3 O 7,} 99%, MW = 294.10), sodium acetate unloading hydroxy Russ _{(C 2 H 3 NaO 2,} MW = 82.03), ethyleneglycol frozen hydroxy Russ _{(C 2 H 6 O 2,} 99.8%, PEG-diacid (COOH-PEG-COOH, MW = 2000) was purchased from Sigma-Aldrich (St. Louis, Mo., USA) Was purchased from Jenkem Technology Co. (Beijing, China). The nanopure H ₂ O (> 18.0 M?) Was purified with a Milli-Q water purification system.

Example  One. Super magnetic  Method for producing nanocomposite

The superparamagnetic nanocomposite in the form of magnetic nanoclusters of Example 1 (specifically superparamagnetic iron oxide nanocomposite) was synthesized by the method shown in Fig.

1.08 g (0.1 M, 3.996 mmol) of FeCl ₃ .6H ₂ O was dissolved in 40 mL of ethylene glycol and stirred for 30 minutes. Then, 0.8 g (0.034 M, 2.720 mmol) of tri-sodium citrate dehydrate (TSC) was added to the stirred mixture and stirred at 900 rpm for 1 hour. After confirming that the tris-sodium citrate was completely dissolved, 2.4 g (0.731 M, 58.515 mmol) of sodium acetate was added to the mixture and stirred again at 900 rpm for 30 minutes. The mixture thus stirred was placed in a Teflon tube for hydrothermal synthesis. The mixture was sealed in a stainless steel container, and the tube was placed in a hydrothermal synthesizer. The mixture was heated from room temperature to 200 ° C at a rate of 7 ° C per minute, The reaction was allowed to proceed for 12 hours. After reaching the maximum temperature (200 ° C), the internal pressure in the sealed synthesis tube was maintained at 1.5 to 2.5 bar.

After the hydrolytic synthesis, the supernatant was removed by magnetic trapping, and the synthesized particles were washed five times with 30 mL of ethanol and five times with deionized water, To prepare a magnetic nanocomposite. The magnetic collection was performed by placing a sample on a Neodymium permanent magnet, collecting the particles, and removing the supernatant. The synthesized particles can also be separated by centrifugation.

As shown in Fig. 1, tris- group) ^Fe--sodium citrate dihydrate carboxylate (COO ^-) groups of the molecule is stable (that is, the surface of Tris-sodium citrate, di-carboxylate (COO molecules of hydrate (COO ^- ) groups of tris-sodium citrate dihydrate molecules are formed by the formation of magnetic nano-crystals, that is, magnetite nanocrystals, which are chemisorbed (anchoring) And stabilized by forming an electrostatic repulsion. On the other hand, in order to reduce the high surface energy of the magnetic nanocrystals, the surface tension acts in the direction of cohesion with each other. A uniform magnitude of superparamagnetic nanocomposite is formed by balancing the electrostatic repulsion and the surface tension.

Example  2. Super magnetic  Method for producing nanocomposite

The superparamagnetic nanocomposite in the form of magnetic nanoclusters of Example 1 (specifically superparamagnetic iron oxide nanocomposite) was synthesized by the method shown in Fig.

2.16 g (0.2 M, 7.991 mmol) of FeCl ₃ .6H ₂ O was dissolved in 40 mL of ethylene glycol and stirred for 30 minutes. Then, 0.02 g (0.25 mM, 0.01 mmol) of PEG-diacid (polyethylene glycol diacid having a molecular weight of 2000) of 0.8 g (0.034 M, 2.720 mmol) was added to the stirred mixture and stirred for another hour. After confirming that the PEG-diacid was completely dissolved, 2.4 g (0.731 M, 58.515 mmol) of sodium acetate was added to the mixture and stirred for another 30 minutes. The mixture thus stirred was placed in a Teflon tube for hydrothermal synthesis. The mixture was sealed in a stainless steel container, and the tube was placed in a hydrothermal synthesizer. The mixture was heated from room temperature to 200 ° C at a rate of 7 ° C per minute, The reaction was allowed to proceed for 12 hours. After reaching the maximum temperature (200 ° C), the internal pressure in the sealed synthesis tube was maintained at 1.5 to 2.5 bar.

After the hydrolytic synthesis, the supernatant was removed by magnetic trapping, and the synthesized particles were washed 5 times with 30 mL of ethanol and 5 times with deionized water, Nanoparticles were prepared. The magnetic collection was performed by placing a sample on a Neodymium permanent magnet, collecting the particles, and removing the supernatant. The synthesized particles can also be separated by centrifugation.

As shown in FIG. 1, the carboxylate (COO ^- ) group of the PEG-diacid molecule stabilizes the surface (that is, chemisorbing, anchoring, and the like) between the carboxylate (COO ^- ) group of the PEG- ) Magnetized nanocrystals are formed, and magnetite nanocrystals are formed and the carboxylate (COO ^- ) group of the PEG-diacid molecule forms stabilized electrostatically repulsion. On the other hand, in order to reduce the high surface energy of the magnetic nanocrystals, the surface tension acts in the direction of cohesion with each other. A uniform magnitude of superparamagnetic nanocomposite is formed by balancing the electrostatic repulsion and the surface tension.

Experimental Example  One. Super magnetic  Physicochemical Properties of Nanocomposites

1-1. Scanning Electron Microscope ( SEM , Scanning Electron Microscope) Super magnetic  Observing the shape of nanocomposites

The size and shape of the super-magnetic nanocomposites of Examples 1 and 2 were observed through a scanning electron microscope (SEM) (S-4700, Hitachi, Tokyo, Japan) Respectively.

FIG. 2 shows the results of observation of the super-magnetic nanocomposites of Example 1 and Example 2 by scanning electron microscopy.

As shown in FIG. 2, in the scanning electron microscope image, the agglomerated structure of the nanocrystals was observed on the surface of the cluster, and the particle sizes of the super-magnetic nanocomposites were 305.9 ± 24.7 nm for Example 1 and 241.7 ± 20.1 nm for Example 2 Of the population.

As can be seen from the scanning electron microscopic photographs of the super-magnetic nanocomposites of Examples 1 and 2, it was confirmed that the relative standard deviation of the distribution of super-magnetic nanocomposites was within 15% .

From these results, it was confirmed that the super-magnetic nanocomposites of Examples 1 and 2 had uniform size and particle size distribution.

1-2. Super magnetic  Size, distribution and surface zeta potential of nanocomposites

The size, distribution and surface zeta potential of the super-magnetic nanocomposites of Examples 1 and 2 were measured using zeta sider (Malvern, model Nano ZS) using dynamic light scattering particle size analysis.

The PDI (PDI = (standard deviation of granularity / average value of granularity) ² } and the surface zeta potential of the super-magnetic nanocomposites of Examples 1 and 2, (3 repeated measurements), and the results are shown in Tables 1-2 and 3-6.

Specifically, Tables 1 and 2 show the average hydrodynamic size, PDI, and surface zeta potential of the super-magnetic nanocomposites of Examples 1 and 2, respectively. FIG. 3 shows the result of measurement of the size of the super-magnetic nanocomposite of Example 1. FIG. 4 shows the result of measurement of the zeta potential of the super-magnetic nanocomposite of Example 1. FIG. FIG. 6 shows the results of measurement of the zeta potential of the super-magnetic nanocomposite of Example 2. FIG.

 [Table 1]

[Table 2]

The measured PDIs of the super-magnetic nanocomposites of Examples 1 and 2 are, according to the numerical value, 0 to 0.1, a nearly monodisperse sample, a mid-range polydisperse of 0.1 to 0.7, ) And > 0.7 sedimentation. The PDI values of Examples 1 and 2 are 0.073 and 0.104, which is less than 0.1 and about 0.1, indicating that the PDI values of Examples 1 and 2 are very good monodisperse . In other words, it was confirmed that the super-magnetic nanocomposites of Examples 1 and 2 had a uniform size and particle size distribution.

In addition, the zeta potentials of the super-magnetic nanocomposites of Examples 1 and 2 correspond to zeta potentials in the range of -15.1 mV and +25.3 mV, respectively, in the range of ± 10-30 mV due to the electrostatic repulsion, It can be judged as a range that can be well dispersed. The super-magnetic nanocomposites of Examples 1 and 2 were stabilized by a carboxylate (COO ^- ) group, so that zeta potentials in the range of ± 10-30 mV were confirmed to be excellent in aqueous solution dispersibility.

1-3. Super magnetic  Magnetic properties of nanocomposites

In order to confirm whether the super-magnetic nanocomposites of Examples 1 and 2 according to the present invention have superior separability to have high magnetization and high magnetic susceptibility to be used for magnetic separation, a superconducting quantum interference device And the magnetic properties were measured. The results are shown in FIGS. 7 and 8. FIG. FIGS. 7 and 8 show the magnetic hysteresis loop of the super-magnetic nanocomposite, and show the result of measuring the magnetic property.

As shown in FIGS. 7 and 8, the super-magnetic nanocomposites of Examples 1 and 2 exhibited superphase magnetic properties at a temperature of 300 K, 76 emu / g for Example 1, 87 emu / g for Example 2 The saturation magnetization values of the samples were shown.

In the case of ferromagnets, it is not suitable for application to magnetic nanoparticle-based separation technology because the residual magnetization is so high that it is strongly influenced by the external magnetic field so that the agglomeration of the particles can occur strongly. However, it is generally difficult to maintain the superparamagnetic properties of magnetic materials at room temperature. In particular, the superparamagnetic-ferromagnetism transition of the magnetic and magnetic complexes of any structure depends on how well the single domains are controlled. . Although the super-magnetic nanocomposite of Examples 1 and 2 according to the present invention has clusters of tens of thousands of super-magnetic nano-crystal units, the final cohesion of the super-magnetic nanocomposite of 200-300 nm has a ferromagnetism transition (Superparamagnetism) without maintaining the superparamagnetism.

In addition, it was confirmed that the super-magnetic nanocomposite of Examples 1 and 2 according to the present invention had a high magnetization and an excellent separation force for magnetic separation.

Experimental Example  2. Super magnetic  Nanocomposite Self Resolution  analysis

2-1. Super magnetic  Using nanocomposites whole blood  (Red blood cells in whole blood) RBCs , Red blood cells) Capture  Experiment

In order to confirm the magnetic separability of the supernatant magnetic nanocomposite of Example 1, only an erythrocyte was collected in the whole blood and the separation efficiency was confirmed.

An anti-RBC antibody (Fitzgerald, Human RBC antibody, Cat # 20R-RR006) was coated on the surface of the super-magnetic nanocomposite of Example 1 in which magnetic nano- 1-Ethyl-3- (3-dimethylaminopropyl) -carbodiimide) / Sulfo-NHS (N-hydroxysulfosuccinimide) mediated covalent bond induction. First, 10 mg / mL of EDC and 50 μL of sulfo-NHS were added to 10 mg / 500 μL of super-magnetic nanocomposite particles, followed by mixing at room temperature for 15 minutes. Then, 1 mg of anti-RBC antibody was mixed with each other, mixed at room temperature for 2 hours, and washed five times with PBS (phosphate-buffered saline) buffer (pH 7.4). A super-magnetic nanocomposite functionalized with an anti-red blood cell antibody was mixed with 25 μL of a PBS-buffer (pH 7.4) of 0.5 mg / 25 μL in a whole blood sample, followed by reaction at room temperature for 5 minutes, The superfine magnetic nanocomposites were separated. Thereafter, the number of red blood cells was observed by observing the supernatant fluid and the supernatant liquid, which were functionalized with the collected anti-red blood cell antibody, by microscopic measurement, and the resolution (%) of red blood cells was calculated according to the following equation Is shown in Fig.

FIG. 9 is a microscope image of a tube photograph taken before and after collection of erythrocytes, and a captured super-magnetic nanocomposite and supernatant.

[Equation 1]

Resolution of red blood cells (%) = number of collected red blood cells / (number of collected red blood cells + number of collected red blood cells) X 100

As shown in Fig. 9, in the left tube photograph in which the whole blood sample and the super-magnetic nanocomposite functionalized with the anti-red blood cell antibody are mixed, erythrocytes are specifically captured and react with the antibody on the surface of the super-magnetic nanocomposite, When only the particles were separated, it was confirmed that the whole blood of red color was seen to be transparent because it was collected by the super magnetic nanocomposite. Observation of the superparamagnetic nanocomposite bound to the erythrocytes by the captured red blood cells and antibody revealed that the erythrocyte (light color) and the super-magnetic nanocomposite (relatively dark black color) It was confirmed that only the red blood cells were not observed. On the other hand, the microscope image of the supernatant contrastively confirmed that the morphology of erythrocytes was not observed.

The erythrocyte separability of the super-magnetic nanocomposite calculated by the formula (1) was 99.5%. As a result, it was confirmed that only the target red blood cells can be specifically captured and rapidly separated even in the whole blood in which non-target immunoglobulin is present in an excess amount of 5 minutes using a magnetic particle.

Claims (20)

Mixing an iron precursor, a solvent, a stabilizer and a reducing agent;
Synthesizing a super-magnetic nanocomposite in nanoclustering form by hydrothermally synthesizing the mixed solution in the mixing step at a temperature of 150 to 300 ° C and a pressure of 1.5 to 10 bar; And
Separating the synthesized super-magnetic nanocomposite,
The stabilizer is a compound having a carboxyl group,
Wherein the superparamagnetic nanocomposite comprises magnetic nano-crystals having a diameter of more than 0 to 10 nm and a surface stabilized by a carboxylate (COO ^- ) group, wherein a plurality of magnetic nanocrystals aggregate and have a diameter of 100 nm to 450 nm A method for preparing a super-magnetic nanocomposite having nanoclusters and having hydrophilic properties dispersed in an aqueous solution. The method of claim 1, further comprising washing the separated superparamagnetic nanocomposite with a polar solvent. The method of claim 1, wherein the mixed solution is subjected to hydrothermal synthesis at a temperature of 200 to 240 ° C and a pressure of 1.5 to 6 bar to synthesize a nanoclustered super-magnetic nanocomposite Wherein the magnetic nanocomposite has a thickness of 10 to 100 nm. The method of claim 1, wherein the iron precursor is iron chloride hexahydrate _{(FeCl 3 · 6H 2 O)} , iron chloride (II), ferric chloride tetrahydrate (II), iron (III) chloride and iron nitrate obtain hydrate (Fe (NO _{3) 3} - > 9H2O). ≪ / _RTI > The method of claim 1, wherein the solvent is selected from the group consisting of ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, dipropylene glycol, and glycerol. delete The composition of claim 1, wherein the stabilizer is selected from the group consisting of tris-sodium citrate dihydrate (HOC (COONa) (CH ₂ COONa) ₂ .2H ₂ O; C ₆ H ₅ Na ₃ O ₇ and dicarboxylic poly (Ethylene glycol). ≪ / RTI > The method of claim 1, wherein the reducing agent is selected from the group consisting of sodium acetate, sodium acrylate, urea, sodium formate, and ammonium acetate. The method of claim 1, wherein the iron precursor and the solvent are mixed in a molar ratio of 1: 10 to 1: 300. The method of claim 1, wherein the iron precursor and the stabilizer are mixed in a molar ratio of 1: 0.0000013 to 1: 1. The method of producing a super-magnetic nanocomposite according to claim 1, wherein the iron precursor and the reducing agent are mixed in a molar ratio of 1: 1 to 1:20. The method of claim 2, wherein the polar solvent is selected from the group consisting of ethanol, water, methanol, acetone, liquid ammonia, ethyl acetate, ether, tetrahydrofuran, potassium hydroxide, sodium hydroxide, and dichloromethane How to. The method of claim 2, wherein the washing of the separated super-magnetic nanocomposite with a polar solvent comprises washing the separated super-magnetic nanocomposite with an ethanol solvent, and washing the super-magnetic nanocomposite washed with the ethanol solvent And washing the resultant nanocrystals with a solvent. The method according to claim 1, wherein the super-magnetic nanocomposite is prepared by adjusting the dispersibility in an aqueous solution by the carboxylate (COO ^- ) group of the stabilizer. delete A magnetic nanocrystal having a diameter of more than 0 nm and having a surface stabilized by a carboxylate (COO-) group, which is produced by the method of any one of claims 1 to 5 and 7 to 14, And having a nanoclustered shape with a diameter of 100 nm to 450 nm and dispersed in an aqueous solution, wherein the magnetic nano-crystals are aggregated with a plurality of magnetic nanocrystals. delete delete The super-magnetic nanocomposite according to claim 16, wherein the magnetic nano-crystal is Fe ₃ O ₄ having a diameter of more than 0 and 10 nm or less. Has a diameter of more than 0 to 10nm or less carboxylate (COO ^-) groups The Fe ₃ O surface is stabilized by ₄ comprises a magnetic nanocrystalline, the plurality of magnetic nano-crystal aggregation is 100 nm to 450 nm of the nano diameter Superparamagnetic nanocomposite with clustered morphology and hydrophilicity dispersed in aqueous solution.

Images